水牛FADS2基因的电子克隆及序列分析

doi:10.16431/j.cnki.1671-7236.2017.10.001

Abstract

Abstract:

This study was aimed to clone the buffalo Δ6-fatty acid desaturases (FADS2) gene using in-Silico cloning and analyze its genetic struction with bioinformatics, which provide a foundation for investigating the milk performance in buffaloes. Primers were designed according the sequence of FADS2 gene in dairy cow (GenBank accession No.:NM_001083444.1). The FADS2 gene was amplified by RT-PCR, and its sequence was analyzed by bioinformatics. Sequence analysis revealed that the buffalo FADS2 gene had 36 600 bp in length and consisted of 12 exons and 11 introns, containing an open reading frame (ORF) of 1 335 bp which encoding 444 amino acids. Sequence homology analysis indicated that the buffalo FADS2 protein gene showed 98.88%, 98.88%, 89.66%, 90.79%, 90.85%, 92.35% and 87.11% identity with that of Bos mutus, Bos taurus, Homo sapiens, Sus scrofa, Oryctolagus cuniculus, Orcinus orca and Rattus norvegicus, respectively. Protein prediction analysis showed that the molecular weight and isoelectric point (pI) of buffalo FADS2 were 52.51 ku and 8.75, respectively, and the FADS2 protein was weak alkali and the hydrophobicity protein without signal peptide. Phylogenetic tree analysis showed that FADS2 gene was highly conserved in different species and evolutionary processes,buffalo was close to Bos mutus and Bos taurus, and was far from Rattus norvegicus. This study suggested that FADS2 gene was successfully cloned in buffalo, which laid a foundation for clarifying the mechanism of milk performance in buffalo.

Key words: buffalo; FADS2 gene; in-Silico cloning; bioinformatics analysis

CLC Number:

MA Xiao-ya, PANG Chun-ying, LIANG Sha-sha, LU Xing-rong, ZHU Peng, DUAN An-qin, LIANG Xian-wei, DENG Ting-xian. Cloning and Bioinformatics Analysis of FADS2 Gene in Buffalo[J]. , 2017, 44(10): 2829-2836.

References

[1] 胡皝,萧浪涛. 生物信息学在新基因全长cDNA电子克隆中的应用[J]. 生物技术通报,2007, 18(4):94-96.
[2] 孙淼,赵茂林. 利用表达序列标签电子克隆cDNA全序列的策略[J]. 生物技术通报,2010, 21(1):49-52.
[3] 卢冉. 鸡Δ6脂肪酸脱氢酶基因启动子区域多态性及基因时空表达的研究[D].郑州:河南农业大学,2011.
[4] 康相涛. 固始鸡-安卡鸡资源群构建及能量与脂肪代谢相关基因遗传变异的研究[D].兰州:甘肃农业大学,2009.
[5] Cho H P, Nakamura M T, Clarke S D. Cloning, expression, and nutritional regulation of the mammalian Delta-6 desaturase[J]. J Biol Chem, 1999, 274(1):471-477.
[6] Osman R H, Liu L, Xia L L, et al. Fads1 and 2 are promoted to meet instant need for long-chain polyunsaturated fatty acids in goose fatty liver[J]. Mol Cell Biochem, 2016, 418(7):103-117.
[7] 丁珍. 乳母乳汁多不饱和脂肪酸水平与FADS2基因多态性的关系研究[D].长春:吉林大学,2016.
[8] Ibeagha-Awemu1 E M, Akwanji K A, Beaudoin F, et al. Association between variants of FADS genes and omega-3 and omega-6 milk fatty acids of Canadian Holstein cows[J]. BMC Genetics, 2014, 15:25.
[9] 徐晨希,王梦琦,朱小瑞,等.中国荷斯坦牛FADS2基因3'端SNP突变对乳中脂肪酸组成的影响[J]. 中国农业科学,2016, 49(11):2194-2202.
[10] Nakayama K, Bayasgalan T, Tazoe F, et al. A single nucleotide polymorphism in the FADS1/FADS2 gene is associated with plasma lipid profiles in two genetically similar Asian ethnic groups with distinctive differences in lifestyle[J]. Human Genetics, 2010, 127:685-690.
[11] 邓廷贤,庞春英,陈明棠,等. 水牛信号转导子和转录激活子1基因电子克隆及序列分析[J]. 中国畜牧兽医,2014, 41(8):9-13.
[12] 刘仲娜,柴志欣,曾贤彬,等. 牦牛CAPN2基因的电子克隆及生物信息学分析[J]. 生物技术,2013, 23(1):42-45.
[13] Aki T, Shimada Y, Inagaki K, et al. Molecular cloning and functional characterization of rat delta-6 fatty acid desaturase[J]. Biochem Biophys Res Commun, 1999, 255(3):575-579.
[14] Marquardt A, Stohr H, White K, et al. Characterization cDNA cloning, genomic structure, and chromosomal localization of three members of the human fatty acid desaturase family[J]. Genomics, 2000, 66(2):175-183.
[15] Green C D, Ozguden-Akkoc C G, Wang Y, et al. Role of fatty acid elongases in determination of de novosynthesized monounsaturated fatty acid species[J]. J Lipid Res, 2010, 51:1871-1877.
[16] 朱士康. 鸡FADS2基因5'端SNP鉴定及启动子功能分析[D]. 郑州:河南农业大学,2014.
[17] Ashwell M S, Heyen D W, Sonstegard T S, et al. Detection of quantitative trait loci affecting milk production, health, and reproductive traits in Holstein cattle[J]. Journal of Dairy Science, 2004, 87(2):468-475.
[18] Alim M A, Wang P, Wu X P, et al. Effect of FASN gene on milk yield and milk composition in the Chinese Holstein dairy population[J]. Anim Genet, 2014, 45(1):111-113.
[19] Tanaka T, Shen J, Abecasis G R, et al. Genome-wide association study of plasma polyunsaturated fatty acids in the In CHIANTI study[J]. PLoS Genet, 2009, 5(1):e1000338.
[20] Nwankwo J O, Spector A A, Domann F E. A nucleotide insertion in the transcriptional regulatory region of FADS2 gives rise to human fatty acid delta-6-desaturase deficiency[J]. J Lipid Res, 2003, 44:2311-2319.
[21] Cobanoglu O, Zaitoun I, Chang Y M, et al. Effects of the signal transducer and activator of transcription 1(STAT1) gene on milk production traits in Holstein dairy cattle[J]. Journal of Dairy Science, 2006, 89(11):4433-4437.
[22] 洪雪莹. 鸡FADS2基因3'UTR区多态性检测及其与经济性状关联性分析[D]. 郑州:河南农业大学,2012.

Cloning and Bioinformatics Analysis of FADS2 Gene in Buffalo

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	XU Yuanyuan, ZHENG Haiying, YANG Chunyan, DENG Tingxian, HUANG Chenqian, FENG Chao, LU Xingrong, DUAN Anqin, MO Xia, MA Xiaoya, SHANG Jianghua. Effects of Inhibition of BMP/Smad Signaling Pathway on the Growth and Steroid Hormone Synthesis of Buffalo Ovarian Granulosa Cells [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2354-2362.
[2]	XIAO Peng, SHANG Jianghua, YANG Chunyan, LI Mengqi, DUAN Anqin, MA Xiaoya, FENG Chao, HUANG Chenqian, ZHANG Bo, ZHOU Jinchen, WEI Kelong, ZHENG Wei, ZHENG Haiying. Effects of Alpha-linolenic Acid on Buffalo Ovarian Granulosa Cells Cultured in vitro [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2380-2387.
[3]	LIU Jialong, ZHANG Yiwen, ZHANG Yueran, SUN Qijuan, CHEN Jian, HE Lei, JIA Yanyan, DING Ke, YU Zuhua. Cloning,Prokaryotic Expression and Bioinformatics Analysis of ELAVL1 Gene in Chicken [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1807-1817.
[4]	ZHUO Na, ZHU Zhongwu, LI Jing, HU Shixiang, WANG Jianchang, LIN Hua, CHEN Chaolin, HAN Diangang, AI Jun, CHEN Peifu. Bioinformatics Analysis and Eukaryotic Expression of ORF2 Protein of Swine Hepatitis E Virus [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1959-1970.
[5]	FAN Bingfeng, LI Wen, LIU Lixiang, ZHAO Xiangyuan, SHAO Jing, LIN Lefeng, SUN Huimin, ZHANG Xulin, XU Baozeng. Cloning and Bioinformatics Analysis of AANAT Gene in Mink [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1307-1318.
[6]	JIA Xinyue, MA Jing, ZHANG Yanyan, MA Xun, WANG Zhengrong, BO Xinwen. Bioinformatics Analysis and Prokaryotic Expression of EGR-07243 and EGR-07243 Genes of Echinococcus granulosus [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 847-858.
[7]	WEI Yirong, ZHENG Zihua, ZHANG Sanbao, SONG Ying, JIANG Haiyu, SUN Wenyue, CHENG Pengfei, LIU Yufan, ZOU Jianwei, HUANG Yanna, PAN Yan, JIANG Qinyang. Cloning,Bioinformatics Analysis and Tissue Expression of UCP1 Gene in Nubian Goats [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 440-450.
[8]	TANG Jingyu, DU Hanyu, MENG Chunchun, LIU Guangqing. Cloning,Bioinformatics Analysis and Subcellular Localization of Interferon-stimulated Gene 15 in Goats [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(8): 2843-2854.
[9]	FU Yongzhi, MA Xiaojun, WANG Maohan, LI Zhe, CHEN Zhiyuan, QUAN Wei, ZHAO Han, SU Yongmei, WU Jianhua, ZHU Wenjin. Bioinformatics Analysis and Tissue Expression of MSTN Gene in Yangyuan Donkeys [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(8): 2910-2919.
[10]	ZOU Jianwei, ZOU Juhong, SHEN Yujian, WEI Yi, ZHANG Sanbao, LIAN Zitong, XU Jianjian, SONG Ying, HUANG Yanna, WEI Yingming, JIANG Qinyang, ZHENG Zihua. Bioinformatics Analysis, Construction of Eukaryotic Expression Vector and Expression of LMCD1 Gene in Nubian Goats [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(6): 2033-2042.
[11]	ZHANG Limeng, LIU Aiju, LI Runting, LI Yuhua, LI Lin, NIE Xiaoning, WANG Linqing, CHEN Longxin. Cloning,Bioinformatics and Tissue Expression Analysis of CREBRF Gene in Ovis aries [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(5): 1599-1609.
[12]	LEI Xing, LI Tianfeng, LI Jiahao, MAN Xin, BAI Suying. Bioinformatics Analysis and Expression of Elovl1 Gene in Muskrat Scent Gland [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(4): 1201-1212.
[13]	SHEN Xiang, CHEN Linghui, YAO Jingjie, WANG Sanhu, BAI Yueyu, ZHANG Xiaojian. Cloning,Bioinformatics and Expression Analysis of Bovine CCR7 Gene [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(4): 1244-1252.
[14]	ZHANG Mingliang, GENG Yachun, LIAN Kaiqi, MA Lei, CHANG Ying, WANG Shuangshan, ZHANG Fuliang. Cloning,Bioinformatics Analysis and Truncated Fragment Expression for Truncated M Gene of Porcine Epidemic Diarrhea Virus [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(4): 1479-1487.
[15]	SHI Yuzhu, LUO Rutang, CHEN Chao, WU Xu, LI Ang. Cloning, Bioinformatics and Tissue Expression Analysis of GDF9 Genes in Muscovy Duck [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(3): 817-829.